Briefly, 1 × Probes Master, 200 nM of each primer,
100 nM Universal ProbeLibrary probe, and 2 μl diluted cDNA template were added to each reaction in a total volume of 20 μl. The protocol consisted of an initial denaturation step at 95°C for 10 min, followed by 40 cycles of amplification and quantification at 95°C for 15 s, 60°C for 10 s, and 72°C for 10 s, and was finally cooled at 40°C. The transcript amounts were estimated from the respective standard curves and normalized to the GAPDH transcript amount determined in corresponding samples. Reactions were run in duplicate. Statistical analysis Results are presented as mean ± SEM. Differences of mean expression levels between groups were compared with the student t-test or Welch’s t-test. Associations were assessed by Pearson’s correlation coefficient test or
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